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This fully updated collection includes chapters on a wide array of techniques that are vital for advancing our understanding of the physiological and pathological effects of estrogen, mediated by estrogen receptor (ER ) and estrogen receptor beta (ERbeta) as well as the non-genomic membrane-bound G-protein coupled estrogen receptor (GPER/GPR30). The protocols range from standard methods and important laboratory workhorses such as immunoblot and RT-qPCR, to newer technologies such as PET imaging, reporter animal models, polysome fractionation, and many others, involving a broad range of tissue types to demonstrate the variety of estrogen receptor effects. Written for this highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative andpractical, Estrogen Receptors: Methods and Protocols, Second Edition serves as an ideal guide to researchers working to explore the vibrancy and excitement of the field of estrogen receptors and estrogen actions.
Sommario
Antibody Validation for Estrogen Receptor Beta.- Immunoblot Detection of the Phosphorylation of the Estrogen Receptor a as an Outcome of GPR30/GPER1 Activation.- Immunoprecipitation Analyses of Estrogen Receptor Phosphorylated at Serine 216 in the Mouse Liver.- Immunofluorescent Verification of Silencing Estrogen Receptor with siRNA in the Intact Rodent Brain.- Detection and Functional Analysis of Estrogen Receptor Phosphorylated at Serine 216 in Mouse Neutrophils.- Quantification of Protein Expression by Proximity Ligation Assay in the Nonhuman Primate in Response to Estrogen.- In Situ Hybridization of Estrogen Receptors and beta in the Human Testis.- Extraction of RNA and Analysis of Estrogen-Responsive Genes by RT-qPCR.- PET Imaging of Estrogen Receptors Using 18F-Based Radioligands.- The Use of ERE-Luc Reporter Mice to Monitor Estrogen Receptor Transcriptional Activity in a Spatio-Temporal Dimension.- Use of Reporter Genes to Analyze Estrogen Response: The Transgenic Zebrafish Model.- Assessing Estrogenic Activity of Classical Estrogen Receptor Binding Compounds.- An Optimized ChIP-Seq Protocol to Determine Chromatin Binding of Estrogen Receptor Beta.- Polysome-Fractionation for Transcriptome-Wide Studies of mRNA Translation.- Anota2seq Analysis for Transcriptome-Wide Studies of mRNA Translation.- Whole-Genome Genotyping Using DNA Microarrays for Population Genetics.- Proteomics Analysis of the Estrogen Effects in the Rat Uterus Using Gel-LC and Tandem Mass Spectrometry Approaches.- Expression Profiles of Estrogen-Regulated MicroRNAs in Cancer Cells.- Molecular Cloning and Purification of the Protein Lysine Methyltransferase SMYD2 and Its Co-Crystallization with a Target Peptide from Estrogen Receptor Alpha.- Molecular Design, Synthesis, and Evaluation of SNIPER(ER) that Induces Targeted Protein Degradation of ER .- Assessment of Metabolic Regulation by Estrogen Receptors.- RNA-Seq Experiment and Data Analysis.
